Methods for using rebaudioside c as a flavor enhancer

ABSTRACT

The present invention is directed to the use of one or more rebaudioside C polymorphs, or stereoisomers thereof, to enhance the sweet taste of Luo Han Guo or Luo Han Guo in combination with one or more of steviol-based glycosides or rebaudioside A.

BACKGROUND OF THE INVENTION

The sweet diterpene glycosides of Stevia have been characterized, andeight sweet glycosides of steviol have been identified. These glycosidesaccumulate in Stevia leaves where they may attain from 10 to 20% of theleaf weight. On a dry weight basis, a typical profile for the four majorglycosides found in the leaves of Stevia includes 0.3% dulcoside, 0.6%rebaudioside C, 3.8% rebaudioside A and 9.1% stevioside. Otherglycosides identified within Stevia include rebaudiosides B, D, and E,and dulcosides A and B. Out of the four major diterpene glycosideflavorings present in Stevia leaves only two (stevioside andrebaudioside A) have physical and sensory properties that are wellcharacterized. Stevioside is known to be 110 to 270 times sweeter thansucrose, rebaudioside A 150 to 320 times sweeter than sucrose,rebaudioside D 200 to 250 times sweeter than sucrose, rebaudioside C 40to 60 times sweeter than sucrose, and dulcoside A 30 times sweeter thansucrose.

Of the diterpene glycosides found in Stevia extracts, rebaudioside A isknown to have the least aftertaste. This aftertaste is described by manyas bitter and licorice-like, and is present in all current Steviaextracts.

Rebaudioside A has been tested in mixtures with other flavorings, suchas fructose, glucose and sucrose, at sweetness intensities equivalent to3% (w/v-%) , 5% (w/v-%) and 7% (w/v-%) sucrose to determine the presenceand degree of synergism in these mixtures (Schiffmann et al., BrainResearch Bulletin 38:105-120 (1995)). According to the results,rebaudioside A appears to have an additive effect in mixtures withfructose and glucose, but a synergistic effect in binary mixtures withsucrose at sweetness intensities equivalent to 3% (w/v-%) sucrose. Atsweetness intensities equivalent to 5% (w/v-%) sucrose, rebaudioside Ahad an additive effect in mixtures with fructose, glucose and sucrose.At sweetness intensities equivalent to 7% (w/v-%) sucrose, rebaudiosideA had an additive effect with a mixture with sucrose, but a suppressiveeffect with mixtures with glucose and fructose. In fact, no flavoringcombinations were synergistic at sweetness intensities equivalent to the7% (w/v-%) sucrose level.

Rebaudioside A has also been tested in ternary mixtures with otherflavorings, such as sucrose, and artificial sweeteners, such as alitame,neohesperidin dihydrochalcone, aspartame, and Na-cyclamate (Schiffmannet al., Chem. Senses 25:131-140 (2000)).

U.S. Pat. No. 4,612,942 mentions that diterpene glycosides can modify orenhance flavor characteristics, such as sweet, when the amount ofditerpene glycoside added is less than the sweetness threshold level ofthe diterpene glycoside in the orally consumable composition. However,no consumable composition for enhancing sweet flavor containingrebaudioside A in combination with rebaudioside C and/or dulcoside A,where the amount of rebaudioside A is less than or equal to the amountof each of rebaudioside C or dulcoside A, is described nor how thesweetness intensity of the consumable composition plays a role in thesweetness enhancing effect of a diterpene glycoside. Further, noconsumable composition for enhancing sweet flavor containingrebaudioside D in combination with rebaudioside C and/or dulcoside A isdescribed.

U.S. Patent Application Publication No. 2009/0162484 A1 describesbeverage products comprising water and a non-sweetening amount of atleast one potent natural sweetener. Examples of such potent naturalsweeteners are described to be one or more of the steviosides,rebaudiosides and related compounds suitable for sweetening. Thepublication does not describe any beverage composition according to thepresent invention.

U.S. Patent Application Publication No. 2009/0162487 A1 describesbeverage products comprising a non-sweetening amount of rebaudioside Aand a sweetening amount of a sweetener other than rebaudioside A.Examples of flavorings other than rebaudioside A are described to benutritive natural flavorings, such as sucrose, glucose, or fructose.However, the publication does not describe any beverage compositionaccording to the present invention.

A need exists for more potent sweet taste enhancers that can effectivelyenhance the sweet taste of a carbohydrate flavoring without exhibitingan off-taste, such as a bitter aftertaste. In particular, a need existsin the art for a method of enhancing the sweetness of consumables thatare already very sweet, i.e., that have a sweetness intensity equivalentto from about 5% (w/v-%) to about 12% (w/v-%) sucrose solution.

BREIF SUMMARY OF THE INVENTION

The present invention is related to the use of at least one rebaudiosideC polymorph, or stereoisomer thereof, for enhancing the sweet taste ofLuo Han Guo or a combination of Luo Han Guo and Steviol-basedglycosides, rebaudioside A, or a combination thereof. A consumable and amethod of decreasing the amount of a flavoring in a consumable are alsoprovided. In particular embodiments, the rebaudioside C polymorph iscrystalline Form I and the at least one rebaudioside C polymorph, or astereoisomer thereof, is present at a concentration of from about 100 μMto about 600 μM. In other embodiments, the at least one rebaudioside Cpolymorph, or a stereoisomer thereof, is administered in a consumablesuch as a food product, a pharmaceutical composition, a dietarysupplement, a nutraceutical, a dental hygienic composition or a cosmeticproduct.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to the use of rebaudioside C (Reb C) forenhancing the sweet taste of flavorings including (a) Luo Han Guo, or(b) a combination of Luo Han Guo and (i) steviol-based glycosides, (ii)rebaudioside A (Reb A), or (iii) steviol-based glycosides and Reb A. Inother embodiments, Reb C and Luo Han Guo are used in combination withammoniated glycyrrhizin (MAGNASWEET), Neohespherdin Dihydrochalcone(NHDC) and/or Thaumatin.

Luo Han Guo is a known sweetener composed of mogrosides obtained fromthe fruit of the perennial vine Siraitia grosvenorii. See U.S. Pat. No.5,433,965. Reb A, 13-[(2-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid β-D-glucopyranosyl ester C.A.S. number58543-16-1) is a diterpenoid glycoside purified or isolated from Steviarebaudiana.

As used herein, steviol-based glycosides are plant extracts containingsteviol glycosides, that may or may not be modified, e.g., with aglucosyl transferase. For example, a steviol-based glycoside can includesteviol glycosides extracted from the Stevia plant, which have beentreated with an enzyme that transfers glucose units from starch to thesteviol glycosides thereby resulting in glucosylated steviol glycosides(see, e.g., WO 2012/129451). Glucosylated steviol glycosides includeNatural Sweet Flavor #2 (NSF02) produced from PURECIRCLE (Oak Brook,Ill.). Another example of steviol-based glycosides is an extract fromRubus sp. (Chaturvedula, et al. (2012) Org. Chem. Curr. Res. 1:1; U.S.Pat. No. 6,228,996).

Reb C,13-[(2-O-α-L-rhamnopyranosyl-3-O-β-D-glucopyranosyl-β-D-glucopyranosyl)oxy]kaur-16-en-18-oic acid β-D-glucopyranosyl ester (C.A.S. number63550-99-2; hereinafter also “Reb C”), is a diterpenoid glycoside. Reb Ccan be used in a purified or isolated form in the present invention, oras an extract from Stevia rebaudiana. Reb C can be prepared by methodsknown in the art, such as that described in U.S. Pat. No. 4,361,697,incorporated herein by reference in its entirety. In one embodiment, RebC is extracted from Stevia rebaudiana, wherein the extract has less than10%, preferably less than 5%, and more preferably less than 3%impurities (i.e., compounds other than Reb C) other than water. Inanother embodiment, one or more polymorphs of Reb C are used. Inspecific embodiments, Reb C crystalline Form I is used in the methods ofthis invention. Crystalline Form I is described in PCT/US2010/049763.

Reb C of the present invention may contain one or more asymmetriccenters and may thus give rise to enantiomers, diastereomers, and otherstereoisomeric forms. The present invention is meant to encompass theuses of all such possible forms, as well as their racemic and resolvedforms and mixtures thereof. The individual enantiomers may be separatedaccording to methods known to those of ordinary skill in the art in viewof the present disclosure. All tautomers are intended to be encompassedby the present invention as well.

As used herein, the term “stereoisomers” is a general term for allisomers of individual molecules that differ only in the orientation oftheir atoms in space. It includes enantiomers and isomers of compoundswith more than one chiral center that are not mirror images of oneanother (diastereomers).

The term “chiral center” refers to a carbon atom to which four differentgroups are attached.

The terms “enantiomer” and “enantiomeric” refer to a molecule thatcannot be superimposed on its mirror image and hence is optically activewherein the enantiomer rotates the plane of polarized light in onedirection and its mirror image compound rotates the plane of polarizedlight in the opposite direction.

The term “racemic” refers to a mixture of equal parts of enantiomers andwhich mixture is optically inactive.

The term “resolution” refers to the separation or concentration ordepletion of one of the two enantiomeric forms of a molecule.

As used herein, the term “sweetness intensity” refers to the relativestrength of sweet sensation as observed or experienced by an individual,e.g., a human, or a degree or amount of sweetness detected by a taster,for example on the scale from 0 (none) to 8 (very strong) used insensory evaluations according to the procedure described in AmericanSociety for Testing Materials, Special Technical Publication-434:“Manual on Sensory Testing Methods,” ASTM International, WestConshohocken, Pa. (1996).

As used herein, the phrase “sweet taste enhancing effect” means that theeffect of Reb C in combination with a flavoring, is such that thesensory perception of the sweet flavor is potentiated in a more thanadditive manner, i.e., synergistically. As used herein, the term“sweetness enhancer” is understood to include at least compositionscapable of enhancing or intensifying the perception of sweet taste offlavoring compositions or flavored compositions. As used herein, theterm “sweetness enhancer” is synonymous with the terms “sweet tastepotentiator,” “sweetness potentiator,” “sweetness amplifier,” and“sweetness intensifier.” Generally, the sweetness enhancers providedherein may enhance or potentiate the sweet taste of a flavoring withoutproviding any noticeable sweet taste by themselves at acceptable uselevels; however, the sweetness enhancers may themselves provide sweettaste at concentrations above a sweetness threshold level. As usedherein, the term “sweetness detection threshold level” is understood toinclude at least the concentration at which the sweetness or off-tasteof an entity is perceptible. The sweetness threshold level varies fordifferent entities, and may be varied with respect to the individual,e.g., the human, perceiving the sweetness.

As used herein, the term “off-taste” refers to an amount or degree oftaste that is not characteristically or usually found in a consumable.For example, an off-taste is an undesirable taste of a sweetenedconsumable to the consumers, such as, a bitter taste, a licorice-liketaste, a metallic taste, an aversive taste, a nasty taste, an astringenttaste, a delayed sweetness onset, and a lingering sweet aftertaste, andthe like.

As used herein in connection with a measured quantity, “about” refers tothe normal variations in that measured quantity, as expected by theskilled artisan making the measurement and exercising a level of carecommensurate with the objective of measurement and the precision of themeasuring equipment.

One or more Reb C polymorphs, or stereoisomers thereof, can be used inconsumables, e.g., in food products, pharmaceuticals, dietarysupplements, nutraceuticals, dental hygienic compositions, or otherproducts as sweetness enhancers, which retain a desired sweetness butcontain lower amounts of the flavoring Luo Han Guo or Luo Han Guo incombination with one or more of steviol-based glycosides or Reb A. Inone embodiment, the present invention provides a consumable, comprisingan effective amount of a combination of Reb C and flavoring in a reducedamount in order to achieve the same level of sweetness when thecarbohydrate flavoring is used alone in the traditional amount. In oneembodiment, the present invention provides a consumable, comprising aneffective amount of Reb C and Luo Han Guo or Luo Han Guo in combinationwith one or more of steviol-based glycosides or Reb A in a reducedamount in order to achieve the same level of sweetness when thecarbohydrate flavoring is used alone in the traditional amount.

In the present invention, one or more Reb C polymorphs, or stereoisomersthereof, is used in an amount effective to enhance the sweetness of LuoHan Guo or Luo Han Guo in combination with one or more of steviol-basedglycosides or Reb A without exhibiting an off-taste. In one embodiment,Reb C is present in an amount less than or equal to the flavoring. Inanother embodiment, the amount of Reb C is greater than or equal to theamount of the flavoring. Any amount of Reb C that provides the desireddegree of sweetness enhancement can be used.

In one embodiment of the present invention, a Reb C polymorph orstereoisomer, is used at a concentration of from about 100 μM to about600 μM. In some embodiments, Reb C is present in the consumable of thepresent invention at a concentration of from about 100 μM to about 350μM. In another embodiment, Reb C is present in the consumable of thepresent invention at a concentration of from about 250 μM to about 350μM. In one embodiment, Reb C is present in the consumable of the presentinvention at a concentration of about 250 μM or about 300 μM or more ofReb C. In one embodiment, Reb C is present in the consumable of thepresent invention at a concentration of about 100 μM, about 150 μM,about 160 μM, about 170 μM, about 180 μM, about 190 μM, about 200 μM,about 210 μM, about 220 μM, about 230 μM, about 240 μM, about 250 μM,about 260 μM, about 270 μM, about 280 μM, about 290 μM, about 300 μM,about 310 μM, about 320 μM, about 330 μM, about 340 μM, or about 350 μM.

In one embodiment, the ratio of one or more Reb C polymorphs, orstereoisomers thereof, to the flavoring is approximately from 1:2, 1:5,1:20, 1:30, 1:40, 1:50, 1:100, 1:150 to 1:200 in a solid consumable. Inone embodiment, the consumable of the present invention contains about0.1 to 0.5 g, preferably about 0.3 g, of one or more Reb C polymorphs,or stereoisomers thereof, for every 50 to 100 g of the flavoring. In oneembodiment, the consumable of the present invention contains about 0.03to 0.15 g of one or more Reb C polymorphs, or stereoisomers thereof, forevery 50 to 100 g of a flavoring.

According to the present invention, one or more Reb C polymorphs, orstereoisomers thereof, act synergistically with the flavoring Luo HanGuo or Luo Han Guo in combination with one or more of steviol-basedglycosides or Reb A, thereby potentiating sweetness intensity even athigh concentrations of the flavoring. In this respect, the amount offlavoring used in a consumable can be reduced. In this respect, theinstant method can be performed such that the amount of flavoringrequired to maintain the desired sweetness of, e.g., a cola beverage isreduced by at least about 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,or 95%, or from about 60% to about 99%, or alternatively from about 20%to about 50%. Thus, in a more specific embodiment, a cola beveragecomprising a flavoring, such as Luo Han Guo or Luo Han Guo incombination with one or more of steviol-based glycosides or Reb A andone or more Reb C polymorphs, or stereoisomers thereof, contains the RebC in an amount sufficient to reduce the amount of sugar required tomaintain the desired sweetness of the beverage by 10%, 20%, 30%, 40%,50%, 60%, 70%, 80%, 90%, or 95%, or from about 60% to about 99%, oralternatively from about 30% to about 70%. Of course, in otherembodiments, the amount of flavoring required can be decreased todiffering extents.

Consumables of the present invention include all food products, dietarysupplements, nutraceuticals, pharmaceutical compositions, dentalhygienic compositions, and cosmetic products. Also, one or moreflavorings other than the flavorings described herein can be present inthe consumables of the present invention, for example, high-intensitysweeteners, such as aspartame, acesulfame potassium, sucralose, andsaccharin or carbohydrate flavorings such as sucrose, fructose, glucose,high fructose corn syrup, xylose, arabinose, rhamnose, erythritol,xylitol, mannitol, sorbitol, or inositol. The flavoring can be presentin the consumable inherently (e.g., in food products containing fruits)or the flavoring is added into the consumable.

The phrase “food product” as used herein includes, but is not limitedto, fruits, vegetables, juices, meat products such as ham, bacon andsausage; egg products, fruit concentrates, gelatins and gelatin-likeproducts such as jams, jellies, preserves, and the like; milk productssuch as ice cream, sour cream, yoghurt, and sherbet; icings, syrupsincluding molasses; corn, wheat, rye, soybean, oat, rice and barleyproducts, cereal products, nut meats and nut products, cakes, cookies,confectionaries such as candies, gums, fruit flavored drops, andchocolates, chewing gum, mints, creams, icing, ice cream, pies andbreads, beverages such as coffee, tea, carbonated soft drinks, such asCOKE® and PEPSI®, non-carbonated soft drinks, juices and other fruitdrinks, sports drinks such as GATORADE®, coffee, teas, iced teas, cola,alcoholic beverages, such as beers, wines and liquors, and KOOL-AID®.Preferably, the food products in which the sweetness of the flavoring isenhanced with one or more polymorphs of Reb C, or a stereoisomerthereof, contains a decreased level of the flavoring. For example, animproved carbonated soft drink can be produced with the same sweetnessas the known carbonated soft drink but with a lower sugar content byadding at least one of Reb C polymorph, or a stereoisomer thereof.

Food products also include condiments such as herbs, spices andseasonings, flavor enhancers, such as monosodium glutamate. A foodproduct also includes prepared packaged products, such as dieteticsweeteners, liquid sweeteners, tabletop flavorings, granulated flavormixes which upon reconstitution with water provide non-carbonateddrinks, instant pudding mixes, instant coffee and tea, coffee whiteners,malted milk mixes, pet foods, livestock feed, tobacco, and materials forbaking applications, such as powdered baking mixes for the preparationof breads, cookies, cakes, pancakes, donuts and the like. Food productsalso include diet or low-calorie food and beverages containing little orno sucrose. Especially preferred food products are carbonated beveragescontaining one or more polymorphs of Reb C, or a stereoisomer thereof.Other examples of food products envisioned in accordance with thepresent invention are described below and throughout the specification.

In another embodiment, the food product is selected from the groupconsisting of fruits, vegetables, juices, meat products such as ham,bacon and sausage; egg products, fruit concentrates, gelatins andgelatin-like products such as jams, jellies, preserves, and the like;milk products such as ice cream, sour cream, yoghurt, and sherbet;icings, syrups including molasses; corn, wheat, rye, soybean, oat, riceand barley products, cereal products, nut meats and nut products, cakes,cookies, confectionaries such as candies, gums, fruit flavored drops,and chocolates, creams, icing, ice cream, pies and breads.

In one embodiment, the invention is directed to a method of decreasingthe amount of a flavoring in a consumable, such as a food product or apharmaceutical composition, to exhibit a given level of sweetness,wherein the method comprises reducing the amount of the flavoring LuoHan Guo or Luo Han Guo in combination with one or more of steviol-basedglycosides or Reb A and adding at least one of Reb C polymorph, or astereoisomer thereof, to the consumable in an amount effective tomaintain the given level of sweetness.

In one embodiment, the food product is a beverage or a drink comprisinga flavoring and one or more Reb C polymporphs, or a stereoisomerthereof. Examples of suitable beverages in which having a sweet taste isdesired include, but are not limited to coffee, teas, such as black tea,green tea, fermented tea, semi-fermented tea, carbonated soft drinks,such as COKE® and PEPSI®, non-carbonated soft drinks, lemonade, juicesand other fruit drinks, sports drinks, such as GATORADE®, iced teas,cola, alcoholic beverages, such as beers, wines and liquors, andKOOL-AID® . In one embodiment, one or more Reb C polymorphs, orstereoisomers thereof, is present at a concentration about 100 μM toabout 600 μM. In one embodiment, the Reb C polymorph or stereoisomer ispresent in the beverage or drink at a concentration of about 100 μM,about 150 μM, about 160 μM, about 170 μM, about 180 μM, about 190 μM,about 200 μM, about 210 μM, about 220 μM, about 230 μM, about 240 μM,about 250 μM, about 260 μM, about 270 μM, about 280 μM, about 290 μM,about 300 μM, about 310 μM, about 320 μM, about 330 μM, about 340 μM, orabout 350 μM. Useful concentrations of Reb C in the beverage or drink ofthe present invention is about 250 μM or about 300 μM, and specifically300 μM.

One embodiment of the invention is directed to a method of enhancing thesweet taste of a cola beverage, such as COKE® or PEPSI®, byadministering to a subject a cola drink, containing Luo Han Guo or LuoHan Guo in combination with one or more of steviol-based glycosides orReb A and at least one Reb C polymorph, or stereoisomer thereof, whereinthe Reb C present in an amount effective to enhance the sweet taste ofthe flavoring without exhibiting any off-taste. In a preferredembodiment, the cola beverage contains a reduced amount of sugar butmaintains substantially the original level of sweet taste. In anotherembodiment, the Reb C is crystalline Form I.

Cola beverages are prepared by mixing cola concentrate with carbonatedwater. Typically about 50 mL of cola concentrate is added per 250 mL ofcarbonated water. Cola concentrate can be prepared by mixing colaflavor, caramel color, and optionally caffeine with water, one or moreflavorings, Reb C, and one or more acid components, such as phosphoricacid or citric acid.

A cola flavor refers to either a natural or artificial flavor. Such colaflavors are commercially available. Commercial cola flavors areavailable, for example, from International Flavor and Fragrances,Dayton, N.J.; Artificial #13573011 and Natural #K3559549. Commercialcola flavors are also available from Tastemaker, Cincinnati, Ohio, andGivaudan Roure, Clifton, N.J.

The acid component refers to an ingredient that contributes sourness tothe beverage and is added to balance the flavor profile. Acids includemalic acid, citric acid, phosphoric acid or combinations thereof.

For example, the cola concentrate can prepared by mixing phosphoric acid(75% Rhone-Poulenc), citric acid (anhydrous, ADM, Decatur, Ill.),caffeine (Mallinckrodt, Paris, Ky.), caramel Color (DS400, Sethness,Chicago, Ill.), cola Flavor (SN018976, International Flavors andFragrances, Dayton, N.J.), sucrose, Reb C, and water. The concentrate isblended until all ingredients are dissolved (30-40 minutes) using amagnetic stirring plate. Fifty milliliters of the concentrate are addedto 250 mL of carbonated water to complete the preparation of the colabeverage. Fifty milliliters of cola concentrate typically contains from0.01 to 5 mL of phosphoric acid, preferably about 0.01-1 mL, 0.1 to 100g of sucrose, preferably about 0.03 g to 0.3 g of Reb C, for every 50 to100 g of sucrose, about 0.001 g to 0.1 g of citric acid, preferablyabout 0.005-0.1 g, 0.001 to 1 g of caffeine, preferably about 0.01 to0.1 g of caffeine, 0.01 to 5 g of caramel flavor, preferably about 0.05to 1 g, 0.001 to about 10 mL of cola flavor, preferably about 0.01 toabout 2 mL.

Food products of the present invention also include animal food productscontaining a flavoring and at least one Reb C polymorph, or stereoisomerthereof, in an amount sufficient to enhance the sweet taste of theflavoring without exhibiting any off-taste. Animal food products arewell known in the art, see, e.g., U.S. Pat. No. 6,403,142, and includedog food, cat food, rabbit food, and the like. The animal food productalso include food products useful for feeding livestock, such as cattle,bison, pigs, chicken, and the like. In another embodiment, the animalfood product of the present invention is a solid hypoallergenic petfood, comprising a component that contains protein or protein fragmentswherein all of said component is partially hydrolyzed and furthercomprises a Reb C polymorph, or stereoisomers thereof.

In one embodiment, the consumable is a pharmaceutical compositioncontaining a flavoring and a Reb C polymorph, or a stereoisomer thereof.These pharmaceutical compositions may be used to formulatepharmaceutical drugs containing one or more active agents that exert abiological effect other than sweetness enhancement. The pharmaceuticalcomposition preferably further comprises one or more active agents thatexert a biological effect. Such active agents include pharmaceutical andbiological agents that have an activity other than taste enhancement.Such active agents are well known in the art. See, e.g., The Physician'sDesk Reference. Such compositions can be prepared according toprocedures known in the art, for example, as described in Remington'sPharmaceutical Sciences, Mack Publishing Co., Easton, Pa., USA. In oneembodiment, such an active agent includes bronchodilators, anorexiants,antihistamines, nutritional supplements, laxatives, analgesics,anesthetics, antacids, H₂-receptor antagonists, anticholinergics,antidiarrheals, demulcents, antitussives, antinauseants, antimicrobials,antibacterials, antifungals, antivirals, expectorants, anti-inflammatoryagents, antipyretics, and mixtures thereof. In one embodiment, theactive agent is selected from the group consisting of antipyretics andanalgesics, e.g., ibuprofen, acetaminophen, or aspirin; laxatives, e.g.,phenolphthalein dioctyl sodium sulfosuccinate; appetite depressants,e.g., amphetamines, phenylpropanolamine, phenylpropanolaminehydrochloride, or caffeine; antacidics, e.g., calcium carbonate;antiasthmatics, e.g., theophylline; antidiuretics, e.g., diphenoxylatehydrochloride; agents active against flatulence, e.g., simethecon;migraine agents, e.g., ergotaminetartrate; psychopharmacological agents,e.g., haloperidol; spasmolytics or sedatives, e.g., phenobarbitol;antihyperkinetics, e.g., methyldopa or methylphenidate; tranquilizers,e.g., benzodiazepines, hydroxinmeprobramates or phenothiazines;antihistaminics, e.g., astemizol, chloropheniramine maleate, pyridaminemaleate, doxlamine succinate, bromopheniramine maleate, phenyltoloxaminecitrate, chlorocyclizine hydrochloride, pheniramine maleate, andphenindamine tartrate; decongestants, e.g., phenylpropanolaminehydrochloride, phenylephrine hydrochloride, pseudoephedrinehydrochloride, pseudoephedrine sulfate, phenylpropanolamine bitartrate,and ephedrine; beta-receptor blockers, e.g., propanolol; agents foralcohol withdrawal, e.g., disulfiram; antitussives, e.g., benzocaine,dextromethorphan, dextromethorphan hydrobromide, noscapine,carbetapentane citrate, and chlophedianol hydrochloride; fluorinesupplements, e.g., sodium fluoride; local antibiotics, e.g.,tetracycline or cleocine; corticosteroid supplements, e.g., prednisoneor prednisolone; agents against goiter formation, e.g., colchicine orallopurinol; antiepileptics, e.g., phenytoine sodium; agents againstdehydration, e.g., electrolyte supplements; antiseptics, e.g.,cetylpyridinium chloride; NSAIDs, e.g., acetaminophen, ibuprofen,naproxen, or salts thereof; gastrointestinal active agents, e.g.,loperamide and famotidine; various alkaloids, e.g., codeine phosphate,codeine sulfate, or morphine; supplements for trace elements, e.g.,sodium chloride, zinc chloride, calcium carbonate, magnesium oxide, andother alkali metal salts and alkali earth metal salts; vitamins;ion-exchange resins, e.g., cholestyramine; cholesterol-depressant andlipid-lowering substances; antiarrhythmics, e.g., N-acetylprocainamide;and expectorants, e.g., guaifenesin.

Active substances which have a particularly unpleasant taste includeantibacterial agents such as ciprofloxacin, ofloxacin, and pefloxacin;antiepileptics such as zonisamide; macrolide antibiotics such aserythromycin; beta-lactam antibiotics such as penicillins andcephalosporins; psychotropic active substances such as chlorpromazine;active substances such as sulpyrine; and agents active against ulcers,such as cimetidine. In another embodiment, the pharmaceuticalcomposition of the present invention comprises at least one amino acidselected from the group consisting of glycine, L-alanine, L-arginine,L-aspartic acid, L-cystine, L-glutamic acid, L-glutamine, L-histidine,L-isoleucine, L-leucine, L-lysine, L-methionine, L-ornithine,L-phenylalanine, L-proline, L-serine, L-threonine, L-tryptophan,L-tyrosine, L-valine, creatine, and mixtures thereof.

The pharmaceutical compositions of the present invention areadministered to a subject in any form suitable to achieve their intendedpurpose. Preferably, however, the composition is one which can beadministered buccally or orally. Alternatively, the pharmaceuticalcomposition may be an oral or nasal spray. The subject is any animal,such as a human, although the invention is not intended to be solimited. Other suitable animals include canines, felines, dogs, cats,livestock, horses, cattle, sheep, and the like. A veterinarycomposition, as used herein, refers to a pharmaceutical composition thatsuitable for non-human animals. Such veterinary compositions are knownin the art.

In another embodiment, the pharmaceutical composition is a liquid dosageform for oral administration, including pharmaceutically acceptableemulsions, solutions, suspensions, syrups, and elixirs. In addition tothe active compounds, the liquid dosage forms may contain inert diluentscommonly used in the art such as, for example, water or other solvents,solubilizing agents and emulsifiers such as ethyl alcohol, isopropylalcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzylbenzoate, propylene glycol, 1,3-butylene glycol, dimethyl formamide,oils (in particular, cottonseed, groundnut, corn, germ, olive, castor,and sesame oils), glycerol, tetrahydrofurfuryl alcohol, polyethyleneglycols and fatty acid esters of sorbitan, and mixtures thereof.Suspensions, in addition to the active compounds, may contain suspendingagents as, for example, ethoxylated isostearyl alcohols, polyoxyethylenesorbitol and sorbitan esters, microcrystalline cellulose, aluminummetahydroxide, bentonite, agar-agar, and tragacanth, and mixturesthereof.

The pharmaceutical composition of the present invention can be in theform of a chewable tablet. Chewable tablets are known in the art. See,e.g., U.S. Pat. Nos. 4,684,534 and 6,060,078, each of which isincorporated by reference in its entirety. Any kind of medicament may becontained in the chewable tablet, preferably a medicament of bittertaste, natural plant extracts or other organic compounds. Morepreferably, vitamins such as vitamin A, vitamin B, vitamin B₁, vitaminB₂, vitamin B₆, vitamin C, vitamin E and vitamin K; natural plantextracts such as Sohgunjung-tang extracts, Sipchundaebo-tang extractsand Eleutherococcus senticosus extracts; organic compounds such asdimenhydrinate, meclazine, acetaminophen, aspirin, phenylpropanolamine,and cetylpyridinium chloride; or gastrointestinal agents such as driedaluminum hydroxide gel, domperidone, soluble azulene, L-glutamine andhydrotalcite may be contained in the core.

The pharmaceutical composition of the present invention can be an orallydisintegrating composition. Orally disintegrating tablets are known inthe art. See, e.g., U.S. Pat. Nos. 6,368,625 and 6,316,029, each ofwhich is hereby incorporated by reference in its entirety.

The pharmaceutical composition of the present invention can be a nasalcomposition, comprising a flavoring, a Reb C polymorph, or stereoisomersthereof. Nasal sprays are known in the art. See, e.g., U.S. Pat. No.6,187,332. Addition of Reb C to a nasal spray can reduce the experienceof an unpleasant taste associated with the composition of the nasalspray.

The pharmaceutical composition of the present invention can be a soliddosage form, comprising a flavoring and a Reb C polymorph, orstereoisomers thereof, and a water and/or saliva activated effervescentgranule, such as one having a controllable rate of effervescence. Theeffervescent composition may further comprise a pharmaceutically activecompound. Effervescent pharmaceutical compositions are known in the art.See, e.g., U.S. Pat. No. 6,649,186, which is incorporated by referencein its entirety. The effervescent composition can be used inpharmaceutical, veterinary, horticultural, household, food, culinary,pesticidal, agricultural, cosmetic, herbicidal, industrial, cleansing,confectionery and flavoring applications. Formulations incorporating theeffervescent composition comprising a Reb C polymorph, or stereoisomersthereof, can further include one or more additional adjuvants and/oractive ingredients which can be chosen from those known in the art,including flavors, diluents, colors, binders, filler, surfactant,disintegrant, stabilizer, compaction vehicles, and non-effervescentdisintegrants.

The pharmaceutical composition can be a film-shaped or wafer-shapedpharmaceutical composition. Such a film-shaped or wafer-shapedpharmaceutical composition can be configured, for example, as quicklydisintegrating administration forms, e.g., administration formsdisintegrating within a period of 1 second up to 3 minutes, or as slowlydisintegrating administration forms, e.g., administration formsdisintegrating within a period of 3 to 15 minutes. The indicateddisintegration times can be set to the above-mentioned ranges by using,for example, matrix-forming polymers which have differentdisintegrating, or solubility, characteristics. Thus, by mixing thecorresponding polymer components, the disintegration time can beadjusted. In addition, disintegrants are known which “draw” water intothe matrix and cause the matrix to burst open from within. As aconsequence, certain embodiments of the invention include suchdisintegrants for the purpose of adjusting the disintegration time.

Suitable are polymers for use in the film-shaped or wafer-shapedpharmaceutical composition include cellulose derivatives, polyvinylalcohol (e.g. MOWIOLT™), polyacrylates, polyvinyl pyrrolidone, celluloseethers, such as ethyl cellulose, as well as polyvinyl alcohol,polyurethane, polymethacrylates, polymethyl methacrylates andderivatives and copolymerisates of the aforementioned polymers.

In certain embodiments, the total thickness of the film-shaped orwafer-shaped pharmaceutical composition according to the invention ispreferably 5 μm up to 10 mm, preferably 30 μm to 2 mm, and withparticular preference 0.1 mm to 1 mm. The pharmaceutical preparationsmay be round, oval, elliptic, triangular, quadrangular or polygonalshape, but they may also have any rounded shape.

In one embodiment, the pharmaceutical composition can be a gum baseformulation comprising a medicament or agent, a flavoring and a Reb Cpolymorph, or stereoisomer thereof, in a coating that surrounds the gumbase formulation. Preferably, the coating comprises at least 50% byweight of the entire product. As the center is chewed, the medicament oragent is released into the saliva. For example, U.S. Pat. No. 6,773,716,which is incorporated herein by reference in its entirety, discloses asuitable medicament or agent contained in a coating that surrounds a gumbase formulation. It has been found that with respect to certainmedicaments or agents that may have an astringent or bitter taste thatby adding a sweet taste enhancing agent to the formulation, that a muchmore palatable formulation, including the medicament, can be provided.In this regard, even though the medicament in, for example, its powderform may be bitter or have an offensive taste, the matrix used as thecoating of the present invention, including the enhancing agent, willafford a product having acceptable medicinal properties.

The pharmaceutical composition of the present invention can be in theform of an aerosol. The aerosol composition may further comprisepharmaceutically active agent. Aerosol compositions are known in theart. See, e.g., U.S. Pat. No. 5,011,678, which is hereby incorporated byreference in its entirety. As a nonlimiting example, an aerosolcomposition according to the present invention may comprise a medicallyeffective amount of a pharmaceutically active substance, one or moreflavorings, a Reb C polymorph, or stereoisomer thereof, and abiocompatible propellant, such as a (hydro/fluoro)carbon propellant.

In one embodiment of the present invention, the pharmaceuticalcomposition is a nutritional composition. Examples of nutritionalcompositions having an undesirable taste include, but are notnecessarily limited to, enteral nutrition products for treatment ofnutritional deficit, trauma, surgery, Crohn's disease, renal disease,hypertension, obesity and the like, to promote athletic performance,muscle enhancement or general well being or inborn errors of metabolismsuch as phenylketonuria. In particular, such nutritional formulationsmay contain one or more amino acids which have a bitter or metallictaste or aftertaste. Such amino acids include, but are not limited to,an essential amino acids selected from the group consisting of L isomersof leucine, isoleucine, histidine, lysine, methionine, phenylalanine,threonine, tryptophan, tyrosine, and valine.

In one embodiment, the sweet taste of the pharmaceutical composition ornutritional composition of the present invention is being enhanced by aReb C polymorph, or stereoisomer thereof, by at least about 10%, 20%,30%, 40%, 50%, 60%, 70%, 80%, 90%, or 95%, or from about 60% to about99%, or alternatively from about 20% to about 50%.

In one embodiment, the consumable of the present invention is a dentalhygienic composition composed of a flavoring, a Reb C polymorph, or astereoisomer thereof, in an amount sufficient to enhance the sweet tasteof the flavoring without exhibiting any off-taste. Dental hygieniccompositions are known in the art and include, but are not necessarilylimited to, toothpaste, mouthwash, plaque rinse, dental floss, dentalpain relievers (such as ANBESOL™) , and the like.

In another embodiment, the consumable of the present invention is acosmetic product containing a flavoring, a Reb C polymorph, or astereoisomer thereof. For example, but not by way of limitation, thecosmetic product can be a face cream, lipstick, lip gloss, and the like.Other suitable compositions of the invention include lip balm, such asCHAPSTICK® or BURT'S BEESWAX® Lip Balm, further containing a Reb Cpolymorph, or a stereoisomer thereof.

The levels of Reb C described herein reflect a creative solution tosweet modulation. As will be appreciated by the skilled artisan, thelevels of Reb C and other sweet materials may vary depending on thelevel of flavoring being reduced as well as the application itself.

In certain embodiments, the preferred levels for Reb C in beverageapplications are 50 ppm, with the range for beverage from 1 ppm to 200ppm and particularly in the range of 50 ppm to 150 ppm.

The other embodiments, particular levels for Reb C in yogurtapplications are 20 ppm, with the range for the application from 10 ppmto 100 ppm and particularly in the range of 10 ppm to 50 ppm.

In still other embodiments, the preferred levels of Reb C in oatmealapplications range from 5 ppm to 400 ppm, 10 ppm to 250 ppm andpreferably from 12 ppm to 100 ppm.

The following examples are illustrative, but not limiting, of thecompounds, compositions, and methods of the present invention. Suitablemodifications and adaptations of the variety of conditions andparameters normally encountered in clinical therapy and which areobvious to those skilled in the art in view of this disclosure arewithin the spirit and scope of the invention.

EXAMPLE 1 Evaluation of Sweetness Intensity

Products are evaluated using a sequential monadic test protocol.Subjects are given a sip (about 2.2 mL from a ca. 1 oz or ca. 30 mLsample) of a control sample into the mouth. The sip is swallowed, thesubject waits for about 15 to 25 seconds, a second sip of the control istaken and swallowed, and the taste is perceived. Thereafter, these stepsare repeated with an experimental sample, and the sweetness can becompared to the control sample. These steps may, for example, berepeated.

EXAMPLE 2 Reb C in Combination with Other Flavorings in Orange StillBeverage

An orange flavored beverage commercially available from InternationalFlavor and Fragrances was prepared (Table 1). The control samplecontained 8 Brix, the measurement of mass ratio of dissolved sugar in aliquid. The sugar used in all experiments was table sugar. an addition,samples containing combinations of rebaudioside C with other known sweetcompounds were evaluated.

TABLE 1 Compound Name Quantity Decanal 0.5% Sinensal 0.2% Ethyl Butyrate0.05%  Orange Oil 1X 5.0% Ethanol 94.25% 

The objective was to determine whether the overall Brix of the beveragecould be lowered 50% and still maintain the same sweetness and temporalprofile of the 8 Brix control. There were ten samples evaluatedincluding; 8 Brix beverage, 4 Brix beverage, four samples of differentcombinations of sweet compounds and four samples of the RP44 added tothe sweet compound combinations. The samples are listed in Table 2.

TABLE 2 Compound Name 1 2 3 4 5 6 7 8 Reb C 150 150 150 150 NSF02 400400 350 350 350 350 350 350 Reb A 20 20 Magnasweet 200 200 NHDC 7.5% 100100 Thaumatin 10 10 *Use Level was ppm

The samples were evaluated by a trained technical panel. The grouprecorded taste comments based on the different combinations of sweetmaterials with and without the addition of rebaudioside C. The tastecomments comparing Test 1 and Test 2 were the following. Test 2 wassweeter and had a more sugary mouthfeel than Test 1. The offnote wasless in Test 2 than Test 1 and slightly sweeter. The comments comparingTest 3 and Test 4 were the following. Test 4 was sweet, bettermouthfeel, cooling, linger, smoother, creamy, enhanced sweetness andmouthfeel. The comments comparing Test 5 and Test 6 were as follows.Test 6 was sweet, clean, and sweeter than Test 5, enhanced mouthfeel andenhanced overall orange flavor. The comments for Test 7 and Test 8 wereas follows. Test 8 has a better mouthfeel, heavy cooked orange, sweet,nice enhancement, sugary mouthfeel, brings the sweetness profile moreupfront.

The samples were also rated based on preference for how similar therebaudioside C samples were to the full sugar control. The ratings scalewas 5 points with 1 being the lowest sweetness or least similar inprofile and 5 the highest sweetness or most similar in profile. The fullsugar was given a rating of 5 and the 50% reduced sugar was given arating of 2. The tests with the rebaudioside C were given the followingratings; Test 2 was a 4, Test 4 was a 2.5, Test 6 was a 3.5 and Test 8was a 3.

In conclusion, the samples containing the rebaudioside C had moremouthfeel and more increased sweetness than the samples without theRP44.

EXAMPLE 3 Rebaudioside C in Combination with Other Flavorings in VanillaFlavored Yogurt

A non-fat commercially available vanilla flavored yogurt was prepared toevaluate flavoring combinations with and without rebaudioside C (Table3). The objective was to determine which yogurt sample gives the closestsweet intensity and temporal profile of the full sugar control. Thecontrol sample contained 8% sugar. The sugar was reduced 40% to 4.8%sugar, and the combinations were added to the base. There were eightcombinations evaluated (Table 4) .

TABLE 3 Compound Name Quantity Vanillin 0.80% Delta Decalactone 0.08%Ethyl Acetate 0.10% Acetyl methyl carbinol 0.08% Furfural 0.06% VanillaExtract 2.00% Ethanol 90.94%

TABLE 4 Compound Name 1 2 3 4 5 6 7 8 Reb C 100 100 80 100 NSF 02 150150 200 200 200 200 150 150 RebA 15 15 Magnasweet 50 50 NHDC 7.5% 80 80Thaumatin 5 5 *Use Level in ppm

The samples were evaluated by a trained technical panel. The paneldescribed the differences between the samples with rebaudioside C to thesamples without rebaudioside C. The comments for the test 1 and test 2were as follows; test 2 had more mouthfeel, sugary profile and enhancedflavor. The comments for test 3 and test 4 were as follows; test 4exhibited enhanced sweetness, enhanced mouthfeel, covered some offnotes, more upfront sweet, sugary mouthfeel, and enhanced flavor. Thecomments for test 5 and test 6 were as follows; test 6 enhancedmouthfeel, smoother, sweeter profile, more upfront sweet, sugarymouthfeel, clean, closer to the control 8% sugar, and enhanced flavor.The comments for test 7 and test 8 were as follows; test 8 enhancedmouthfeel, enhanced sweetness, enhanced vanillin flavor, and wasslightly browner.

In addition, the samples containing the rebaudioside C were ratedagainst the full sugar control. The ratings were based on a 5 pointscale; with 5 being the sweetest and closest in profile and 1 being theleast sweet and similar in profile. The rating for the full sugar samplewas a 5. The other samples rated in the following ascending order. Themost similar was test 4 with a rating of 4.5, test 1 with a rating of 4,test 3 with a rating of 3 and test 2 with a rating of 1.

In conclusion, the samples that had the rebaudioside C performed betterthan the samples without the rebaudioside C in all combinations.

EXAMPLE 4 Reb C in Combination with Luo Han Guo

Cereal, Lemon-Lime-flavored carbonated soft drink (CSD), orange-flavoredbeverage (Orange Still) and yogurt were prepared with combinations ofrebaudioside C, NSF02 and Luo Han Guo and the distance from sugar waspredicted.

TABLE 3 Distance Luo from Consumable Reb C NSF02 Han Guo Sugar* Cereal27% (152 ppm) 68% (380 ppm) 5% (30 ppm) 6.89 Lemon-Lime 4% (12 ppm) 93%(273 ppm) 3% (9 ppm)  1.56 CSD 3% (10 ppm) 94% (275 ppm) 3% (9 ppm) 1.56 36% (121 ppm) 64% (216 ppm) 2.01 Orange 7% (47 ppm) 93% (627 ppm)1.00 Still 2% (9 ppm)  93% (523 ppm) 5% (28 ppm) 0.41 2% (12 ppm) 94%(534 ppm) 4% (25 ppm) 0.42 Yogurt 95% (315 ppm) 5% (16 ppm) 1.91 7% (22ppm) 93% (298 ppm) 2.18 *The smaller the number, the better.

What is claimed is:
 1. A method of enhancing a sweet taste of aflavoring, comprising administering to a subject at least onerebaudioside C (Reb C) polymorph, or a stereoisomer thereof, incombination with a flavoring selected from: (a) Luo Han Guo, or (b) LuoHan Guo in combination with: (i) steviol-based glycosides, (ii)rebaudioside A, or (iii) a combination of (i) and (ii), wherein the RebC is administered in an amount effective to provide a sweet tasteenhancing effect without exhibiting an off-taste.
 2. The method of claim1, further comprising administering ammoniated glycyrrhizin,neohespherdin dihydrochalcone or thaumatin.
 3. The method of claim 1,wherein the rebaudioside C polymorph is crystalline Form I.
 4. Themethod of claim 1, wherein the flavoring and at least one rebaudioside Cpolymorph, or a stereoisomer thereof, is administered in a consumable.5. The method of claim 4, wherein the consumable is a food product, apharmaceutical composition, a dietary supplement, a nutraceutical, adental hygienic composition or a cosmetic product.
 6. The method ofclaim 1, wherein the at least one rebaudioside C polymorph, or astereoisomer thereof, is present at a concentration of from about 100 μMto about 600 μM.
 7. A consumable comprising at least one rebaudioside C(Reb C) polymorph, or a stereoisomer thereof, in combination with aflavoring selected from: (a) Luo Han Guo, or (b) Luo Han Guo incombination with: (i) steviol-based glycosides, (ii) rebaudioside A, or(iii) a combination of (i) and (ii), wherein the Reb C is in an amounteffective to enhance the sweet taste of the flavoring without exhibitingan off-taste.
 8. The consumable of claim 7, further comprising anammoniated glycyrrhizin, neohespherdin dihydrochalcone or thaumatin. 9.The consumable of claim 7, wherein the rebaudioside C polymorph iscrystalline Form I.
 10. The consumable of claim 7, wherein theconsumable is a food product, a pharmaceutical composition, a dietarysupplement, a nutraceutical, a dental hygienic composition, a cosmeticproduct or a tabletop flavoring.
 11. The consumable of claim 7, whereinthe at least one rebaudioside C polymorph, or a stereoisomer thereof, ispresent at a concentration of from about 100 μM to about 600 μM.
 12. Amethod of decreasing the amount of a flavoring in a consumablecomprising adding to a consumable containing a flavoring selected from:(a) Luo Han Guo, or (b) Luo Han Guo in combination with: (i)steviol-based glycosides, (ii) rebaudioside A, or (iii) a combination of(i) and (ii), at least one rebaudioside C (Reb C) polymorph, or astereoisomer thereof, thereby reducing the amount of the flavoringneeded to exhibit a given level of sweetness.
 13. The method of claim12, further comprising adding an ammoniated glycyrrhizin, neohespherdindihydrochalcone or thaumatin.
 14. The method of claim 12, wherein therebaudioside C polymorph is crystalline Form I.
 15. The method of claim12, wherein the consumable is a food product, a pharmaceuticalcomposition, a dietary supplement, a nutraceutical, a dental hygieniccomposition, a cosmetic product or a tabletop flavoring.
 16. The methodof claim 12, wherein the at least one rebaudioside C polymorph, or astereoisomer thereof, is present at a concentration of from about 100 μMto about 600 μM.